EP4344786A1

EP4344786A1 - Biomagnetic separation system with double ring profile

Info

Publication number: EP4344786A1
Application number: EP22382891.4A
Authority: EP
Inventors: Lluis Miquel Martinez Garcia
Original assignee: Sepmag Systems SL
Current assignee: Sepmag Systems SL
Priority date: 2022-09-27
Filing date: 2022-09-27
Publication date: 2024-04-03
Also published as: CN117790108A; JP2024048395A; US20240112839A1

Abstract

Biomagnetic separation system comprising an outer ring and an inner ring of permanent magnets, the outer ring being a quadrupole Halbach cylinder and the inner ring being a Halbach cylinder with N poles. The diameter of the cylinders and other conditions are chosen so that at the inner radius of the inner ring the magnetic field gradient is greater at the vessel wall than the magnetic field gradient of the outer ring alone.

Description

BACKGROUND OF THE INVENTION

Field of the Invention

The present invention relates to the field of magnetic separation of particles. More in particular, the invention refers to a biomagnetic separator for large volumes.

Description of the Related Art

Magnetic separation systems have many applications in the field of medicine, biology and pharmacology. Particular elements of a sample, suspension or solution (for instance some types of antibodies), often need to be separated in order to analyze aspects regarding these elements (like diagnosing an illness). The methods traditionally used to achieve this type of separation of elements, particles or molecules are the method of separation by affinity columns and the centrifugation method.
Another method, whose use has increased in recent years, is a method of separation based on the use of magnetic particles. This method is quick and easy for precise and reliable separation of elements such as, for example, specific proteins, genetic material and biomolecules (see, for example, Z M Saiyed, et al., "Application of Magnetic Techniques in the Field of Drug Discovery and Biomedicine". BioMagnetic Research and Technology 2003, 1:2, published 18 September 2003 (available at http://www.biomagres.com/content/1/1/2 ). The method is based on the use of magnetic particles designed to join to the specific elements that are to be separated from a sample, solution, suspension, etc., in some type of vessel. By applying a magnetic field, the magnetic particles are separated from the rest of the sample or, rather, are concentrated at the walls of the vessel, where they are retained (for example, due to the magnetic field which is applied) while the rest of the sample (or, at least, a substantial part of the rest of the sample) is removed. The retained fraction can subsequently be subjected to a washing process which may include another separation of magnetic particles, etc.
Separators of magnetic particles based on the structure disclosed in U.S. Pat. No. 5,705,064 can generate intense magnetic fields, while separators based on the structure disclosed in US-A-2003/0015474 can generate almost constant magnetic field gradients. These structures are based on the Halbach Theorem, which demonstrates that if the magnetization of an infinite linear magnet magnetized perpendicularly to its axis is rotated around this axis, the magnetic field is constant in module throughout the space and its direction turns in all of the space in the same angle in the direction opposite to rotation (K. Halbach, "Design of permanent multipole magnets with oriented rare earth cobalt material", Nuclear Instruments and Methods Volume 169, ). Using this principle, dipolar sources can be developed which produce uniform fields inside cylindrical cavities (see, for example, H. A. Leupold, "Static Applications" in "Rare Earth Permanent Magnets", J. M. D. Coey (Editor), 1996, pages 401-405). In addition, a near zero magnetic field can be achieved outside the cylinder, something which is advantageous in terms of safety. These structures are also known as "Halbach Cylinders".
The principle can be easily used on multipolar sources, achieving, in the case of four pole sources, a constant gradient. These structures are functional and present, in theory, no major technical problems when small volumes are involved (applied to recipients of volumes in the order of a few ml). The magnetic field gradient generated by the Halbach cylinder of inner radius Ro and external radius R₂ , will generate a constant magnetic field gradient over the magnetic particles, generating a radial movement to the inner walls of a cylindrical vessel of inner radius Zo inserted in a bore coaxial with the cylinder (Z₀ <R₀ ).
Once the magnetic particles are separated (i.e. all of them arrive to their final positions), the suspension liquid is removed. At this point the magnetic field gradient should be strong enough to keep all the magnetic particles retained in the inner walls of the vessel, even when is not liquid, avoiding the loss of magnetic particles and the biomolecules attached to them. For a given volume concentration of particles in the suspension, the surface density of magnetic particles retained in the inner cylindrical wall of the vessel at the end of the separation process will increase linearly with its radius. Then the magnetic field gradient needed to retain the magnetic particles will be higher for larger radius vessels.
However, the magnetic field gradient generated by a quadripolar Halbach cylinder will be ∇B=2*B_r /R₀*(1-R₀ /R₂), where B_r is the remanence of the permanent magnet used. Even in the case of an infinitely high cylinder with R ₂ infinite (R₀ /R ₂->0), the gradient will be inversely proportional to the radius and with a limit ∇B >2*B_r /R₀.
When increasing the radius of the vessel Z₀ for increasing the batch volume of the magnetic separation process, a suspension of magnetic particles will require an increased magnetic field gradient to cope with the larger surface density of magnetic particles at the retention area, while the magnetic field gradient will decrease, limited by the inner radius of the bore (R₀ >Z₀ ). At a critical radius, the gradient generated by the quadripolar Halbach cylinder will be smaller than the value needed for retaining the magnetic particles when the suspension liquid is removed.

SUMMARY OF THE INVENTION

The invention solves the problems above by providing a magnetic separator with an outer ring comprising a quadrupolar Halbach cylinder and an inner ring made of permanent magnets with a particular number of poles and inner and outer radius that depend on the filling factor of the magnets and the radii of the vessel and the outer ring. The inner ring provides a magnetic field gradient at Z₀ which retains the particles and does not compromise the separation capability of the outer ring.
In this way, slipping-down of the separated particles at the inner walls of the vessel is avoided, while maintaining a high productivity of the separation process and reducing the amount of the rare-earth magnets necessary for achieving the sought magnetic field gradient.
More in particular, the biomagnetic separation system of the invention has a double ring profile comprising an outer ring with inner radius R₁ and outer radius R₂ of n₂ >4 permanent magnets of the same geometry and a magnetization progression of ${Δγ}_{2} = 3 {Δθ}_{2}$
where Δθ ₂ Is the angular distance between two consecutive magnets, and an inner ring with outer radius R₁ and inner radius R₀ of n₁ permanent magnets of the same geometry, with n₁ >2N, N being the number of pole pairs, the inner ring magnets having a magnetization progression of $Δγ = (N + 1) Δθ$
where Δθ Is the angular distance between two consecutive segments, where the inner ring is concentric with the outer ring and defines an inner bore for placing a vessel whose inner face is at Z₀ from the geometric center of the rings, the outer ring having a remanence Br₁ and filing factor f₁ the inner ring having a remanence Br₂ and filling factor f₂ and wherein the outer ring and inner ring fulfill the following conditions: ${Nz}^{N - 2} > 2 kf (1 - \frac{R_{0}}{R_{2}})$
$R_{1} > \frac{R_{0}}{\sqrt[N - 1]{1 - \frac{2 kf (1 - \frac{R_{0}}{R_{2}})}{{Nz}^{N - 2}}}}$
with k >1, z being the ratio Z₀ /R₀ and f=(Br₂*f₂)/(Br₁*f₁).
In the particular case that all the magnets used for both rings are the same, and the filling factor is also the same, f=1.
The outer ring can be made however of sub-rings of magnets with different remanence and the filling factor of both rings can be different.

BRIEF DESCRIPTION OF THE DRAWINGS

To complete the description and provide for better understanding of the invention, a set of drawings is provided. Said drawings illustrate a preferred embodiment of the invention, which should not be interpreted as restricting the scope of the invention, but just as an example of how the invention can be carried out.

Figure 1 shows the working principle of a system according to the invention.
Figure 2 shows the description of a double magnetic field gradient separation system according to the invention.
Figure 3 is a graph showing the magnetic field gradient exerted by both rings versus the normalized radius of the device.
Figure 4 shows a double magnetic field gradient separation system according to the invention. In this particular embodiment, the magnets used for the Halbach cylinder (outer and inner rings) have a square cross-section.
Figure 5 shows the magnetic field gradient profile along the radius at θ=0° at the embodiment shown figure 4.

DESCRIPTION OF THE INVENTION

With reference to Figure 1, the working principle of the large-volume magnetic separation of beads/particles is as follows: a vessel containing the suspension is introduced into the separation system (Figure 1 (a)). The particles move radially to the walls of the vessel, dragged by the magnetic field gradient (Figure 1 (b)). When the supernatant/buffer is extracted from the vessel, the particles are retained on the walls of the receptacle thanks to the application of a second magnetic field gradient as will be explained later (Figure 1 (c)).
If the magnetic field gradient would not be strong enough on the walls of the receptacle, the particles would not be fully retained on the walls as in the present invention (Figure 1 (d)). In order to solve this problem, the present invention as shown in Figure 2 proposes a double-ring approach. An outer ring made of a plurality of concentric sub-rings and forming a quadrupole Halbach cylinder (the number of pole pairs, N, is 2) generates a magnetic field with a constant gradient high enough to separate the particles. An inner ring generates a higher polar number field (N>2), with a shorter reach but a higher magnetic field gradient at the retention position Z₀ . The inner ring defines the inner space or bore of the device, in which a vessel containing the suspension is to be placed.
Assuming the height of the ring is larger than its inner radius, for a given radial position Z₀ , a single quadripolar Halbach cylinder with inner radius R₀ (R₀ >Z₀) and external radius R ₂ (R₂ >R₀) would generate a magnetic field gradient of B_r2.f₂ /R₀*(1-R₀ /R₂), where Br₂ is the remanence of the permanent magnets used for building the system and f₂ the filling factor (f₂ =1 when the ring is manufactured by magnets filling all the ring, f₂ <1 if the geometry of the magnets doesn't fill all the space).
For having a magnetic field gradient at Z₀ (k>1) k-times higher than that of the quadrupole Halbach cylinder, the invention provides an inner magnetic ring with inner radius R ₀, and external radius R₁ (same as the inner radius of the outer ring), made of permanent magnets with remanence B_r1 , filling factor f₁ , and its number of pole pairs N fulfilling the condition: ${Nz}^{N - 2} > 2 kf (1 - \frac{R_{0}}{R_{2}})$
where z is the ratio Z₀ /R₀ and f=(B_r2*f₂)/(B_r1f₁), f=1 if the filling factor and magnets remanence are both the same for the inner and outer rings (B_r1 =B_r2, f₁ =f₂ ). The value of the outer radius of the inner ring R₁ (R₀<R₁<R₂ ) should be $R_{1} > \frac{R_{0}}{\sqrt[N - 1]{1 - \frac{2 kf (1 - \frac{R_{0}}{R_{2}})}{{Nz}^{N - 2}}}}$
The factor k is the ratio between the magnetic field gradient necessary for safely retaining the magnetic beads when the suspension liquid is removed, and the magnetic field gradient generated by a quadripolar Halbach Cylinder with inner radius R₀ and outer radius R₂ , filled with permanent magnets with remanence Br₂ and a filling factor f₂ and capable of separating the particles.
All the relations above apply obviously to all cases where the dimensions of the separation device are R₀>Z₀ and the height of the rings, h, greater that the internal diameter of the inner ring (h>2R₀ )
The inner ring should then be manufactured with n₁ segments of permanent magnets with the same geometry, with n₁ > 2N, each one with an angular progression of the magnetization, Δγ, $Δγ = (N + 1) Δθ$
where Δθ Is the angular distance between two consecutive segments (Δθ=2π/n₁).
The outer ring, with inner radius R₁ and outer radius R₂ , should be built with the number of segments n₂ >4. The angular progression of the magnetization should be ${Δγ}_{2} = 3 {Δθ}_{2}$
where Δθ₂ Is the angular distance between two consecutive segments (Δθ₂ =2π/n₂ ).
The resultant double ring device generates a magnetic field gradient larger than the equivalent conventional quadrupolar Halbach cylinder alone at the position Z₀, while the gradient in the inner part of the vessel wall will be 2B_r2.f₂ /R₁*(1-R₁ /R₂).
As shown in Figure 3, the profile of the separation magnetic field gradient G_sep , contrary to that of the retention magnetic field gradient G_ret, is constant in the whole volume of the inner space. The retention magnetic field gradient is noticeable only in the vicinity of the interior walls of the device, that is, close to the vessel's walls.

Example:

A device was built for separating the magnetic beads from a biological suspension contained in a vessel with a diameter 286.5 mm and a wall thickness of 4.1 mm. When the vessel is filled, the height of the liquid is 400 mm. For ensuring that all magnetic beads are retained in the inner walls of the vessel (Z ₀ =139.2 mm) when all the supernatant is extracted, it is necessary a radial magnetic field gradient of at least 15 T/m. In the present example, the outer ring was made of two sub-rings and the magnet's remanence was the same both for the outer and inner ring.
The device is shown in figure 4 and has the following features:

An inner ring with an inner radius of R₀ =150 mm and outer radius R₁ =172 mm is manufactured using 36 magnets of 20x20x400 mm, magnetized along the 20 mm direction, B_r1 =1.32 T, with the center at R=164 mm from the cylindrical axe (the geometrical center of both rings, as they are concentrical) and separated by 10° and with their magnetization direction rotating by 100° between consecutive magnets (number of pole pairs, N=9). The filling factor of this ring is f₁ =0.65.
An outer ring with an inner radius of R ₁=172 mm and outer radius R ₂=282 mm is manufactured with two sub-rings of 40x40x400 mm magnets magnetized along the 40 mm direction, with Br₂ =1.32 T. The first sub-ring has 24 magnets with the center at R=199 mm from the cylindrical axe and separated by 15°, with their magnetization direction rotating by 45° between consecutive magnets. The second sub-ring is composed of 32 magnets with the center at R=232 mm from the cylindrical axe and separated by 11.25° with their magnetization direction rotating by 33.75° between consecutive magnets. The filling factor of the outer ring is f₂ =0.57.

The magnets will be enclosed in an Aluminium frame with an inner diameter of 296 mm, an outer diameter of 568 mm, and a height of 400 mm with the corresponding housing for the magnets. The system will be enclosed with a 10 mm thick top and a bottom cover with the same diameters as the Aluminium frame. The resultant device weighs 405 kg. 308 kg corresponds to the permanent magnets and 97 kg to the Aluminium frame and covers.
As shown in figure 5, the magnetic field gradient generated by the inner ring at Zo is 20.2 T/m and the outer ring generates a constant gradient of 3.4 T/m. The retention gradient at Z₀ is higher than 15 T/m, fulfilling the magnetic field gradient specifications.
As it is used herein, the term "comprises" and derivations thereof (such as "comprising", etc.) should not be understood in an excluding sense, that is, these terms should not be interpreted as excluding the possibility that what is described and defined may include further elements, steps, etc.
On the other hand, the invention is obviously not limited to the specific embodiment(s) described herein, but also encompasses any variations that may be considered by any person skilled in the art (for example, as regards the choice of materials, dimensions, components, configuration, etc.) to be within the general scope of the invention as defined in the claims.

Claims

Biomagnetic separation system with double ring profile comprising an outer ring with inner radius R₁ and outer radius R ₂ of n₂ >4 permanent magnets of the same geometry and a magnetization progression of ${Δγ}_{2} = 3 {Δθ}_{2}$
where Δθ₂ Is the angular distance between two consecutive magnets,
and an inner ring with outer radius R₁ and inner radius R₀ of n₁ permanent magnets of the same geometry, with n₁ >2N, N being the number of pole pairs, the inner ring magnets having a magnetization progression of $Δγ = (N + 1) Δθ$

where Δθ Is the angular distance between two consecutive segments, where the inner ring is concentric with the outer ring and defines an inner bore for placing a vessel whose inner face is at Z₀ from the geometric center of the rings,

the outer ring having a remanence Br₁ and filing factor f₁

the inner ring having a remanence Br₂ and filling factor f₂

and wherein the outer ring and inner ring fulfill the following conditions: ${Nz}^{N - 2} > 2 kf (1 - \frac{R_{0}}{R_{2}})$
$R_{1} > \frac{R_{0}}{\sqrt[N - 1]{1 - \frac{2 kf (1 - \frac{R_{0}}{R_{2}})}{{Nz}^{N - 2}}}}$
With k >1, z being the ratio Z₀ /R₀ and f=(Br₂*f₂)/(Br₁*f₁).
Biomagnetic separation system with double ring profile according to claim 1, where f=1, the filling factor and magnets remanence being the same for the inner and outer rings.